Computational protocol: A fibril-specific, conformation-dependent antibody recognizes a subset of Aβ plaques in Alzheimer disease, Down syndrome and Tg2576 transgenic mouse brain

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Protocol publication

[…] To obtain measures of co-localization between OC-positive immunolabeling and neuritic plaques or diffuse plaques, additional images were analyzed from confocal microscopy experiments. Confocal images captured from three Down syndrome cases and with three controls were used to obtain either neuritic plaque (thioflavin S-positive with OC) or diffuse plaque colocalization (using OC and 6E10 labeled sections in cases that were thioflavin S-negative). One image was captured (using a 20× objective such that 1 pixel = 0.18 μm) for each case for each double labeling experiment. Image J (Rasband, W. S., ImageJ, U. S. National Institutes of Health, Bethesda, MA, USA, http://rsb.info.nih.gov/ij/, 1997–2008) and the JACoP plug in [] was used to obtain Manders’ overlap coefficients, which provides a proportion of the “green signal” coincident with the “red signal”. Images were thresholded prior to quantification by first determining a cut off for the marker (OC or thioflavin or 6E10) that occupied the most area in the image with the second marker automatically set to the same threshold. […]

Pipeline specifications

Software tools ImageJ, JACoP
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Mus musculus, Homo sapiens
Diseases Alzheimer Disease, Dementia, Neurotoxicity Syndromes